Chapter title |
Quantitation of 5-Methyltetrahydrofolate in Cerebrospinal Fluid Using Liquid Chromatography-Electrospray Tandem Mass Spectrometry
|
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Chapter number | 19 |
Book title |
Clinical Applications of Mass Spectrometry in Biomolecular Analysis
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Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3182-8_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3181-1, 978-1-4939-3182-8
|
Authors |
Erland Arning, Teodoro Bottiglieri |
Abstract |
We describe a simple stable isotope dilution method for accurate and precise measurement of cerebrospinal fluid (CSF) 5-methyltetrahydrofolate (5-MTHF) as a clinical diagnostic test. 5-MTHF is the main biologically active form of folic acid and is involved in regulation of homocysteine and DNA synthesis. Measurement of 5-MTHF in CSF provides diagnostic information regarding diseases affecting folate metabolism within the central nervous system, in particular inborn errors of folate metabolism. Determination of 5-MTHF in CSF (50 μL) was performed utilizing high performance liquid chromatography coupled with electrospray positive ionization tandem mass spectrometry (HPLC-ESI-MS/MS). 5-MTHF in CSF is determined by a 1:2 dilution with internal standard (5-MTHF-(13)C5) and injected directly onto the HPLC-ESI-MS/MS system. Each assay is quantified using a five-point standard curve (25-400 nM) and has an analytical measurement range of 3-1000 nM. |
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